Hornschuh Melanie, Haas Vivian, Winkel Paul P, Gökyildirim Mira Y, Mullins Christina S, Wrobel Ida Maria, Manteuffel Christian, Wirthgen Elisa
Department of Pediatrics, Rostock University Medical Center, 18057 Rostock, Germany.
Medical School, Rostock University, 18057 Rostock, Germany.
Biology (Basel). 2022 Oct 27;11(11):1583. doi: 10.3390/biology11111583.
Monocyte-derived macrophages or dendritic cells are of increasing interest for cellular therapeutic products to treat inflammation-related diseases and cancer. However, the isolation method and the culture conditions applied influence the functionality of cells. For some approaches, the adhesion-induced differentiation into macrophages must be prevented to maintain functions attributed to circulating monocytes. The effects of the isolation method on the functionality of non-adherent peripheral monocytes have not yet been investigated.
The present study examines the impact of the isolation method on cell viability, growth, metabolism, inflammation-induced cytokine response, migratory capacity, and adherence of non-adherent human peripheral monocytes. The monocytes were isolated by magnetic sorting using either positive or negative selection and cultured in cell-repellent plates.
The purity and yield of monocytes were higher after positive selection. However, the adherence and migratory capacity, cytokine response, and metabolic activity were decreased compared to negatively selected monocytes. The impaired functionality presented in combination with cell shrinking, thus, indicates the start of cell viability loss. Negatively selected non-adherent monocytes showed no impairment in functionality, and the viability remained high. In conclusion, this approach is better suited for conducting ex vivo modifications of monocytes prior to the intended experimental setup or therapeutic application.
单核细胞衍生的巨噬细胞或树突状细胞在治疗炎症相关疾病和癌症的细胞治疗产品中越来越受到关注。然而，所采用的分离方法和培养条件会影响细胞的功能。对于某些方法，必须防止因黏附诱导分化为巨噬细胞，以维持循环单核细胞所具有的功能。分离方法对非黏附性外周单核细胞功能的影响尚未得到研究。
本研究考察了分离方法对非黏附性人外周单核细胞的细胞活力、生长、代谢、炎症诱导的细胞因子反应、迁移能力和黏附性的影响。通过使用阳性或阴性选择的磁性分选法分离单核细胞，并在细胞排斥板中进行培养。
阳性选择后单核细胞的纯度和产量更高。然而，与阴性选择的单核细胞相比，其黏附性和迁移能力、细胞因子反应及代谢活性均有所降低。功能受损与细胞萎缩同时出现，因此表明细胞活力开始丧失。阴性选择的非黏附性单核细胞在功能上未表现出受损，且活力仍然很高。总之，这种方法更适合在预期的实验设置或治疗应用之前对单核细胞进行体外修饰。
